Main sol. 1053
Internal Solution-ID: S2129
Equipment needed: Filter
Autoclave
Hungate tubes
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NaCl | 20.00 | g | 19.782 | 338.508 | |
| K2HPO4 | 0.09 | g | 0.089 | 0.511 | |
| KH2PO4 | 0.07 | g | 0.069 | 0.509 | |
| CaCl2 | 0.80 | g | 0.791 | 7.13 | |
| NH4Cl | 0.25 | g | 0.247 | 4.623 | |
| NaNO3 | 1.00 | g | 0.989 | 11.637 | |
| MgSO4 x 7 H2O | 4.00 | g | 3.956 | 16.052 | |
| MgCl2 x 6 H2O | 3.00 | g | 2.967 | 14.596 | |
| KCl | 0.33 | g | 0.326 | 4.378 | |
| Fe2(SO4)3 x H2O | 0.01 | g | 0.01 | - | |
| Modified Wolin's mineral solution | 10.00 | ml | - | - | |
| Na2S2O3 x 5 H2O | 1.00 | g | 0.989 | 3.985 | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 4.9e-4 | 0.002 | |
|
Sulfur
(powdered) |
10.00 | g | 9.891 | 308.425 | |
| NaHCO3 | 1.00 | g | 0.989 | 11.774 | |
| Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients (except sulfur, bicarbonate and vitamins), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 6.8 with NaOH and dispense under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials (up to a volume of 20%) that contain already the appropriate amount of sulfur. Autoclave at a temperature of 110°C for 20 min. Add vitamins and bicarbonate to the autoclaved medium from sterile anoxic stock solutions. The solution of vitamins is sterilized by filtration and stored under 100% N2 gas, whereas the solution of bicarbonate is prepared under 80% N2 and 20% CO2 gas mixture and autoclaved. Adjust the pH of the complete medium to 6.5. | |||||
| 2 After inoculation pressurize vessels to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. | |||||
Media with this solution
| 1053 | SULFURIMONAS PARALVINELLA MEDIUM |